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Victor Fries was one of GothCorp's most brilliant scientists and cryogenic researchers. When his wife Nora Fries became infected with a deadly disease, he developed a special freezing cell and planned to cryogenically hold her until a cure was found. However, Fries had been misappropriating GothCorp money, leaving the company in debt, and the experiment unauthorized. When he was interrupted by GothCorp CEO Ferris Boyle, a fight ensued and Fries grabbed a handgun, pointing it at Boyle and his security guards. Fries was then calmly ordered to drop the gun; reasoning that it is not his nature to resort to violence and that a possibility may exist for him to legitimately bankroll Nora's experiment. When Boyle's smooth talk got to Fries as he relaxed and let down his guard, the deceitful CEO kicked the scientist into a lab table full of beakers which exploded. The resulting explosion smashed Fries into his cryogenic freezing tanks and the accident soaked his entire body with the freezing solution and rendered him unable to survive outside of a sub-zero environment. Vowed revenge on those who had wronged him, he became known as "Mr. Freeze" after creating a vacuum tight suit which maintained his body temperature at 50 degrees below zero and tripled his strength as well as building a laser-powered cold gun.

Freeze is introduced in the episode "Heart of Ice", which won an Emmy for Outstanding Writing in an Animated Program. Freeze leads a band of henchmen and makes several raids on the company GothCorp, stealing the parts for a freezing machine he wishes to build and use in his battle against the company. This leads to his first confrontation with Batman, whom he defeats easily and traps in a block of ice. Later on, before another attack on Gothcorp after the machine is complete, Freeze catches Batman in the security room, watching an archive video revealing Freeze's tragic origin. While working for Gothcorp, Victor Fries had been using illegal funding for an experiment to save his dying wife, Nora, by cryogenically freezing her to preserve her until a cure could be found. At the last moment, however, the ruthless Gothcorp CEO, Ferris Boyle, had broken into the lab with guards and kicked Fries into a batch of chemicals, leaving him to die.

Fries survived, but was severely mutated by the substances and transformed into Mr. Freeze. Now, on a vendetta against Boyle, Freeze traps Batman and leaves him in his hideout, confronting Boyle at the Gothcorp Humanitarian Party and freezing him up to the waist. Batman, however, escapes and confronts Freeze at the party, fighting him one-on-one until he breaks Freeze's helmet with a flask of chicken soup. Freeze is defeated, and arrested along with Boyle, as Batman presented evidence of Boyle's crimes to the press. Victor Fries is remanded to Arkham Asylum, but wearing a standard prison uniform as a special cell with ultralow temperatures was made for him.

It should be noted that Batman's sympathies were with Mr Freeze in this episode, especially after he witnessed the accident that spawned Freeze. He even tried to reason with Freeze but was unsuccessful because (as the good Doctor put it) his emotions had been "frozen dead" within him. However, this is disproven at the end of episode as an imprisoned Fries is shown crying that he failed in his mission to avenge his wife.

"I can only beg your forgiveness, and pray you hear me somehow, someplace... someplace where a warm hand waits for mine"

Freeze later appears in "Deep Freeze", where he is sprung out of prison (against his will) and taken to an off-shore city named Oceana by an android owned by theme-park builder Grant Walker, who is fascinated with Freeze and needs his technology to condition his body to become like Freeze's, for his maniacal plan of a Global Deep Freeze to create a new world for his selected residents of Oceana. Freeze, at first, refuses to help Walker, but when Walker shows Freeze his wife, alive but still cryogenically frozen, Freeze accepts. Batman and Robin arrive and tell him that if Freeze helps Walker freeze Gotham, Nora would hate him.

Freeze finally sees the truth, and helps Batman and Robin stop Walker at the last minute. Freeze battles Walker, who has been placed in a cryo-suit identical to Freeze's, and wins by pinning Walker to a wall with his gun. Freeze then proceeds to overload Oceana's power-core, blowing the city to smithereens and warning the inhabitants to flee for their lives. Freeze opts to stay behind and die with his wife, and freezes Robin to stop Batman from following him. Freeze, Nora and Walker disappear in the explosion, but the episode's end reveals that they all survive, temporarily trapped in icebergs.

In Batman & Mr. Freeze: SubZero, Fries settles in the Arctic, accompanied only by his frozen wife, and an Inuit boy, Kunac, and two polar bears he adopted. An expedition submarine compromised the integrity of Fries' cave, causing ice to smash open Nora's capsule. Enraged by the unaware expedition's unintentional mistake, Victor freezes them all, and moves back to Gotham.

While there, he enlists the help of former colleague and ethically-corrupt doctor Gregory Belson. Belson informed Freeze Nora's condition required major organ transplant, and her blood type was rare. He kidnapped Barbara Gordon after Belson found her to be a matching donor for Nora. The operation was to take place on an abandoned oil rig.

Batman and Robin intervene, and the ensuing struggles set the rig ablaze. Belson abandoned Freeze, only to meet his own demise. Freeze suffered a broken leg, and insisted that Batman rescue Nora and Kunac. Batman saves them and Barbara, then went back for Freeze. However, the rig's structure was compromised by the surrounding fire and explosions, and Freeze fell into the sea below. His polar bears rescued him and he later returned to the Arctic, walking with a crutch and a cast on his foot. Peering into a scientific post, he caught the news on television than Nora made a full recovery with the help of research from Wayne Enterprises. Happy tears in his eyes, Fries limped away into the Blizzard.

Freeze returns in The New Batman Adventures episode "Cold Comfort", his new-found joy quickly evaporated. Nora had recovered, but believing her husband dead, remarried and left Gotham. At the same time, Fries' own condition worsened, and by the time kidnapped doctors were able to stabilise him, Fries was reduced to a head in a jar walking on mechanical spider-like legs, interacting with the outside world with a robotic suit. Faced with no probability of assuming a normal life with Nora, Freeze decides to take out his pain and loss on Gotham by freezing everyone and everything in it. Batman thwarted Fries, who went missing afterward. The next time Fries resurfaces would be decades later in the Batman Beyond episode, "Meltdown".

Fries' head was in storage at a facility belonging to Wayne-Powers, kept alive by having machines take care of his vital functions and not having aged a minute. Derek Powers, now Blight, is searching for a cure to his mutation. Dr. Stephanie Lake, a young ambitious researcher, recommends growing a new body for Powers, and transferring his consciousness into the new body. Victor Fries was to be the test subject.

Fries was thrilled to be able to feel again after the procedure, and decides to turnover a new leaf. Bruce Wayne is suspicious of his old foe, and instructs Terry McGinnis, the new Batman, to monitor him. Terry was not so convinced on the old man's suspicion.

Fries' new-found life was short, however, as he inexplicably relapses. At Derek Powers' instructions, Dr. Lake locked Victor in a chamber and increased the heat to weaken him enough to run a biopsy to determine the problem.

Freeze managed to escape, however, and returned with a vengeance in his new cryogenic suit, which he had left in cold storage. For Powers' and Lake's treachery, Mr. Freeze freezes them, despite Lake pleading for mercy. Lake is presumably killed at this point, encased completely in cold ice and no visible breathing holes. Freeze proceeds to destroy the facility. Batman tried to stop him, but Derek Powers, now in his Blight guise, broke out of his ice encasing and intervened, fighting them both.

Freeze finally managed to dispatch of Blight, blowing him out of the building, but his suit was damaged by Blight's radiation blast. Batman pleaded with Freeze to escape, but Freeze was determined to go down with the building, saying to Batman in a sad regretful tone, "Believe me... You're the only one who cares..." Freeze seals himself off. Batman escapes as the building collapses. Freeze is presumably killed in the incident.

Terry later admitted to Bruce he was wrong about Freeze. Bruce replied that they both were.

BTAS revised several smaller Batman villains, but none of them were as successful as Mr. Freeze. Freeze had been introduced in the comics with little depth or detail as one of the many gimmick oriented enemies of Batman. The character was completely altered for the series, which introduced a new sympathetic backstory, his wife Nora, and great moral complexity. The character's cold, stoic demeanor coupled with a barely restrained vindictive fury compared him to Batman himself. This version of Mr Freeze was shown to be very popular with fans, and the comics soon resurrected him, as he was previously killed by the Joker and the comics variation of Mr Freeze adopted this backstory for the character. Even the film Batman and Robin, though featuring a far less serious interpretation of the character, included elements of the backstory, such as Nora's demise and the accident which transformed Victor.